title: Single-step purification of immunoglobulin M on C1q-Sepharose.
abstract: A rapid and simple affinity chromatography method for purifying IgM from myeloma serum and ascites fluid is described. Complement protein C1q is coupled to Sepharose with an efficiency of 35%, giving 1.7 mg of C1q bound/ml of gel. This C1q-Sepharose selectively binds IgM from crude samples at 5 degrees C, with a capacity of 0.4 mg of IgM/ml of gel. The bound IgM may be eluted simply and isocratically by bringing the gel to room temperature for 2 h, or by washing with buffer containing 0.5 M KI. The eluted IgM is highly pure by SDS-PAGE and double immunodiffusion analysis, although IgG may be a potential contaminant. The C1q-Sepharose is stable for at least 18 months.
authors.affiliation: Clinical Immunology Research Centre, University of Sydney, NSW, Australia.
authors.forename: A
authors.initials: A
authors.lastname: Nethery
authors.forename: R L
authors.initials: RL
authors.lastname: Raison
authors.forename: S B
authors.initials: SB
authors.lastname: Easterbrook-Smith
mesh_terms: D002846:Chromatography, Affinity; D015922:Complement C1q / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007075:Immunoglobulin M / Q000302:isolation & purification*; D012685:Sepharose
chemical_list: D007075:Immunoglobulin M; D015922:Complement C1q; D012685:Sepharose